1.Clone and Expression of Loop1 and Loop2 Gene of Hexonof Infectious Canine Hepatitis Virus
Long ZHENG ; Jun-Xia WANG ; Li-Min LI ; Xia ZHANG ; Huan-Ling ZHANG ; Hong-Yu YOU ;
China Biotechnology 2006;0(04):-
The mainly antigenic sites for the adenovirus neutraliation are present on Loop1 and Loop2 of hexon.Majority research were focus in the human adenovirus.Little was known on infectious canine hepatitis virus (ICHV), which was also called canine adenovirus typeⅠ.Here,ICHV (the isolated strain) DNA was isolated and purified from the cultured MDCK cells.The Loop1 and Loop2 fragments were amplified by polymerase chain reaction(PCR) method,and then was connected by ligase T4.The target fragment was then connected with vector pET28a.The nucleotide sequence ecoding Loop1 and Loop2 was determined.The nucleotide sequence identity of Loop1 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 100%, 100% and 83.8%, and the nucleotide sequence identity of Loop2 region between the isolated strain and CLL, RI261 and Toronto A26/61 strains is 88.1% , 88.1% and 99.3%, and amino acid identity is 93.6%, 93.6% and 98.6%.The recombinant Loop protein was expressed in E.coli and was approximately 36kDa in size,and then was purified. Then BALB/c mice were injected subcutaneously in the back and armpit with the recombinant Loop protein.The anti-ICHV antibody titers of immunized serum was tested by indirect ELISA and the titers were up to 1:320.Western blot demonstrated that immunized sera could specifically combine with ICHV. The research laid a foundation for creating new genetic engineering products of infectious canine hepatitis virus.
2.Clinical application of the tracking scan technique in 16-slice spiral CT angiography
Renhua ZHANG ; Bohong DENG ; Jiangtao LONG ; Qiang LI ; You YANG ; Meiying CHEN
Chinese Journal of Medical Imaging Technology 2009;25(7):1296-1299
Objective To explore the application of the tracking scan technique in 16-slice spiral CT angiography, in order to improve the quality of CTA. Methods Three hundred patients who were divided into three groups randomly underwent CTA in tracking, testing and estimation scan respectively with GE LightSpeed 16-slice spiral CT. The data of all patients were transmitted to the workstation (AW4.2) and reconstructed. The quality of all images were evaluated by three experienced doctors with double-blind method and divided into four grades (A, B, C and D) from optimal to poor. Results The rate of grade A, B, C and D was 89.00%, 7.00%, 4.00% and 0 respectively for tracking scan, 70.00%, 11.00%, 15.00% and 4.00% respectively for testing scan, while 61.00%, 13.00%, 21.00% and 5.00% respectively for estimation scan. Conclusion Tracking scan technique is superior to testing and estimation scan in image quality of 16-slice spiral CT angiography.
3.Mutual effect between neuropeptides and inflammatory cytokines in neurogenic SMSCs of human temporomandibular joint.
Zhi-Ming, LIU ; You-Jian, PENG ; Xing, LONG ; Jian, LI ; Jin, KE ; Wei, FANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):602-7
In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.
4.The protective role of angiotensinⅡreceptor inhibitor in ventilator-induced lung injury in rats
Dan FENG ; Shang-Long YAO ; You SHANG ; Qing-Ping WU ; Li-Kui WANG ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To study the protective role of AngiotensinⅡreceptor inhibitor in ventilator-induced lung injury of rats.Method Forty healthy male SD rats were equally divided into four group (A,B,C,D group,n=10).Group A served as control group,group B had low tidal volume (V_T=10 ml/kg) with breathing rate (P)=80/min;group C had high tidal volume (V_T=40 ml/kg) group with breathing rate 80/min;group D had high tidal volume (V_T=40 ml/kg) group with breathing rate 80/min,all rats in group D were pretreated with Losartan.The duration of ventilation in 'all groups was two hours.Rats were sacrificed after experiment finished. The lung lavage liquid and lung tissue were collected and preserved with well established methods.Lung pathological change was observed by microscope;lung cell apoptosis was assessed with TUNEL;the expression of ANGⅡwas assayed with RT-PCR.The measured variables also included total protein,WBC,W/D,MPO. Results In comparison with B group,all variables in group C were significantly increased (P
5.The differential expression of P57kip2 and CDK5 in retinoic acid-induced neural tube defects
Xinjun LI ; Yangyun HAN ; Hong XU ; Zhong YANG ; Yi ZENG ; Zhongshu SUN ; Hongli LI ; Xiaodong LONG ; Chao YOU
Chongqing Medicine 2013;(28):3344-3346
Objective To investigate the differential expression of P57kip2 and CDK5 in neural tube defects t(NTD) from the normal ,and provide the clue for the research of the molecular mechanism of the normal neurula formation .Methods A cDNA mi-croarray containing 1 100 known genes was used to compare differences in P57kip2 and CDK5 gene expression between the normal control group and the retinoic acid(RA)-induced NTD group on embryonic(E) day 9 .5 and 10 .5 .Two differentially expressed genes were randomly selected from the two groups for Northern blotting to verify the results of the cDNA microarray .Results Compared the differences of between P57kip2 and CDK5 in normal and E9 .5 d ,E10 .5 d ,E9 .5 d-NTD ,E10 .5 d-NTD ,P57kip2 and CDK5 expression was significantly up-regulated in the before and after the formation of the normal neurulation ,but them showed a downward trend in retinoic acid (RA)-induced NTD(including two phase E9 .5 d and E10 .5 d) .Conclusion P57kip2 and CDK5 in-volved in the physiological process of NTD ,and provide the useful clue for the research of the molecular mechanism of the normal neurula formation .
6.Change of renin-angiotensin system in cultured mesangial cells by serum from nephrectomized rats feeding with low protein diet and α-keto acid supplement
Ying YIN ; Quan LONG ; Li YOU ; Haiming LI ; Liyin ZHANG ; Yong GU ; Chuanming HAO ; Shantan LIN ; Jing CHEN
Chinese Journal of Nephrology 2011;27(6):435-441
Objective To observe the changes of renin-angiotensin system (RAS) in cultured mesangial cells by serum from 3/4 nephrectomized rats feeding with low protein diet and α-keto acid. Methods Thirty male SD rats received 3/4 nephrectomy (Nx) were placed on 18%normal protein diet (NPD,n=10),6% low protein diet(LPD,n=10) or 5% low protein plus 1%α-keto acid diet (LK,n=10) flor 12 weeks.Ten male SD sham-operated rats fed with 18% normal protein diet were used as control (sham group).In addition,mesangial cells were cultured in sera (10%) collected from above animals treated with or without losartan (0.02 mmol/L)for 48 hours.ELISA was applied to detect the level of Ang II,TGF-β1 and fibronectin (FN) in cell medium.Westem blotting was used to determine the protein level of ATI receptor (AT1R)and real-time PCR was used to detect the mRNA level of AT1R,TGF-β1 and FN. Results (1) Nutritional indices including body weight,total protein and albumin had no significant difference in each group. (2) Serum creatinine and 24 h pruteinuria were significantly inceased in nephrectomized groups compared to sham group(P<0.05,respectively).24 h proteinuria was greatly lower in LK group than that in NPD and LPD groups(P<0.05,respectively).(3)LK greatly decteased the level of Ang II[NPD(12.70±0.12)mg/g protein;sham(8.04±0.62)mg/g protein]in supernatant as well as the protein and mRNA expression of AT1R in cultured mesangial cells (P<0.05).(4)NPD serum directly induced higher secretion[FN:sham(20.58±0.46)g/g protein,NPD (39.84±0.06)g/g protein;TGF-β1:sham(10.12±O.56)mg/g protein,NPD(83.85±3.58)mg/g protein] and mRNA expression of FN and TGF-β1 compared with sham group (P<0.05).LPD decreased these increment (P<0.05) and LK showed stronger inhibitory effect (P<0.05). (5)Losartan application sharply reduced FN and TGF-β1 production both in supematant and in mRNA expression in NPD serum treated cells (P<0.05,respectively). Conclusion Low protein diet with α-keto acids supplement directly inhibits the RAS in mesangial cells which may contribute to its beneficial effect on the kidney.
7.Clinical observation on the efficacy of Endostar combined with platin-based chemotherapy for 55 cases of advanced non-small cell lung cancer
Jianlin LONG ; Lu LI ; Meijuan HUANG ; Li REN ; Mei HOU ; Jin WANG ; Yong XU ; Feng PENG ; You LU
Tumor 2010;(2):156-159
Objective:To observe the efficacy, median progression-free survival (PFS) and adverse reaction induced by rh-endostatin injection (Endostar) plus platin-based chemotherapy for advanced non-small cell lung cancer (NSCLC).Methods:Fifty five histologically or cytologically confirmed advanced NSCLC patients received Endostar combined with platin-based chemotherapy for more than 2 cycles. The evaluated parameters included PFS, response rate (RR), clinical benefit rate (CBR) and adverse reaction. Results:Of the 51 patients who can be evaluated for response, 15 (29.4%) achieved partial response (PR), 27 (52.9%) had stable disease (SD), 9 (17.6%) had progressive disease(PD), no patient had complete response(CR). The overall RR was 29.4% (15/51) and CBR was 82.4% (42/51). The median PFS was 6.3 months. There were no significant differences in the short-term efficacy and PFS between the patients who had different pathological features (P=0.037), those had naive or relapsed diseases (P=0.101), or those received different chemotherapeutic regimens (P=0.232). The total white cells and platelets decreased by 72.7% and 54.5%, respectively. The frequency of grade Ⅲ or Ⅳ neutropenia and thrombocytopenia were 36.4% (20 caces) and 21.8% (12 cases), respectively. Four patients stopped the therapy for adverse reaction. One died of gastrointestinal hemorrhage; one had uncontrolled grade Ⅲ hypertension; one had superventricular arrhythmia; one had grade Ⅳ hepatic dysfunction. Conclusion:The combination of Endostar and platin-based chemotherapy increased the CBR and prolonged the PFS of the patients with advanced NSCLC. The toxicities were tolerable.
8.Testis specific serine/threonine protein kinase 4 (TSSK4) leads to cell apoptosis relying on its kinase activity.
Xiao-li WANG ; You-heng WEI ; Guo-long FU ; Long YU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2015;35(2):235-240
Testis specific serine/threonine protein kinase 4 (TSSK4) belongs to the TSSK family, and its members play an important role in spermatogenesis and/or spermiogenesis. Mouse TSSK4 has been reported to be expressed exclusively in the testis and can maintain its kinase activity through autophosphorylation at Thr-197. However, its biological function remains poorly understood. Here we found that GFP-TSSK4-overexpressed HeLa cells showed apoptotic bodies, indicating TSSK4 can lead to apoptosis in vitro. Furthermore, TSSK4 induced apoptosis in different cell lines including HeLa, Cos-7 and H1299 tested by flow cytometry but not its kinase-dead mutant TSSK4-K54M. TSSK4 knockout mice showed increased testes weight and decreased apoptotic spermatogonia and spermatocytes at 21st day after birth tested by TUNEL technology. So TSSK4 was able to induce cell apoptosis in vitro depending on its kinase activity, which leads to abnormal testes weight and apoptosis, shedding light on its function in the process of spermatogenesis and/or spermiogenesis.
Animals
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Apoptosis
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physiology
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Base Sequence
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Cell Line
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DNA Primers
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Flow Cytometry
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Humans
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In Situ Nick-End Labeling
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Male
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Mice
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Polymerase Chain Reaction
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Protein-Serine-Threonine Kinases
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physiology
9.Comparison of structural characteristics and anticoagulation activity of enoxaparin sodium with different degree of 1,6-anhydro derivatives.
He ZHU ; You-Jing LIÜ ; Xian-Wei HAN ; Shi-Long LIU ; Jie-Jie HAO ; Xiao-Liang ZHAO ; Guang-Li YU
Acta Pharmaceutica Sinica 2014;49(7):1049-1053
The fine structure of enoxaparin sodium samples with different degree of 1,6-anhydro derivatives were analyzed with polyacrylamide gel electrophoresis, high performance liquid chromatography, ultraviolet spectroscopy, infrared spectroscopy and nuclear magnetic resonance spectroscopy. A further study of anticoagulation activity of enoxaparins was performed, including those on their inhibition activities of coagulation factor Xa (FXa) and thrombin (FIIa). The results showed that the anti-FXa and -FIIa activities of enoxaparins with different degree of 1,6-anhydro derivatives (20.0%-39.7%) with similar structure characteristics, had decreasing tendency when the degree of 1,6-anhydro derivatives increased. Especially, the anti-FXa activity was sensitive to the change of the degree of 1,6-anhydro derivatives.
Anticoagulants
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chemistry
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Enoxaparin
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chemistry
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Factor Xa Inhibitors
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chemistry
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Thrombin
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antagonists & inhibitors
10.Applications of the hospital statistics management system.
Hong ZHAI ; Yong REN ; Jing LIU ; You-Zhang LI ; Xiao-Long MA ; Tao-Tao JIAO
Chinese Journal of Medical Instrumentation 2008;32(1):64-69
The Hospital Statistics Management System is built on an Office Automation Platform of Shandong provincial hospital system. Its workflow, role and popedom technologies are used to standardize and optimize the management program of statistics in the total quality control of hospital statistics. The system's applications have combined the office automation platform with the statistics management in a hospital and this provides a practical example of a modern hospital statistics management model.
Hospital Administration
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Hospital Information Systems
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Office Automation
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Statistics as Topic